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. 2000 Jan 18;97(2):710–715. doi: 10.1073/pnas.97.2.710

Figure 3.

Figure 3

Methylation analysis of multiple CpG islands in colorectal adenomas. (A) Methylation of multiple CpG islands detected by methylated CpG island amplification. Methylated CpG island amplification products from colon adenomas (P2–P48) were blotted on a nylon membrane and were hybridized with one of the MINT clones (indicated on the top), as well as a p16 probe. M2, MINT2; M1, MINT1; M12, MINT12; and M31, MINT31. Tumors P46, P47, and P48 showed methylation of multiple loci, indicating the presence of CIMP. Tumor P23 showed methylation of MINT31 only. (B) Methylation analysis of MINT2 by bisulfite-PCR. PCR products were digested with BstUI, which cleaves CGCG sequences that are retained after bisulfite treatment only when both cytosines are methylated. Methylated alleles are shown by arrows. Tumors P46, P47, and P48 are methylated at this locus. No methylated alleles were detected in PCR products from normal colon. RKO is a colon cancer cell line used as a positive control. (C) Methylation analysis of p16 in colorectal adenomas by MSP. U, unmethylated allele-specific primers; M, methylated allele-specific primers. Tumor P28 and P33 showed methylated alleles.